Optimized production of transgenic buffalo embryos and offspring by cytoplasmic zygote injection.

ABSTRACT: BACKGROUND:Cytoplasmic injection of exogenous DNA into zygotes is a promising technique to generate transgenic livestock. However, it is still relatively inefficient and has not yet been demonstrated to work in buffalo. We sought to improve two key technical parameters of the procedure, namely i) how much linear DNA to inject and ii) when to inject it. For this, we introduced a constitutively expressed enhanced green fluorescent protein (EGFP) plasmid into buffalo zygotes. RESULTS:First, we found that the proportion of EGFP-expressing blastocysts derived from zygotes injected with 20 or 50 ng/μL DNA was significantly higher than from those injected with 5 μg/mL. However, 50 ng/μL exogenous DNA compromised blastocyst development compared to non-injected IVF controls. Therefore the highest n

SUBMITTER: Meng F 

PROVIDER: S-EPMC4617447 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature